M-CSF和IL-10对人单核细胞表面分子表达的影响

本文采用流式细胞术检测人外周血单核细胞表面CD14、CD2 3、CD6 4、CD11b、CD18、CD2 9表达 ,研究巨噬细胞集落刺激因子 (M CSF )和白细胞介素 (IL ) 10对单核细胞炎症效应和免疫效应功能的影响。结果显示 ,(1)M CSF诱导单核细胞表面CD14及CD2 3分子的表达 (P <0 0 5 )。IL 10抑制CD2 3的表达 ,促进CD6 4的表达 ,并协同M CSF诱导单核细胞CD14的表达 ,拮抗M CSF对CD2 3的诱导作用。M CSF能协同IL 10对单核细胞CD6 4的诱导作用 ;(2 )M CSF能诱导单核细胞表面CD11b及CD18的表达 (P <0 0 5 )。结论 :M CSF通过促进单核细胞表面CD11b、CD18、CD14、CD2 3的表达及协同促进CD6 4的表达而促进单核细胞粘附、炎性渗出、IgG及IgE依赖的细胞杀伤和吞噬功能 ,促进单核 巨噬细胞在炎症反应、体液免疫应答效应阶段发挥效应细胞功能。IL 10通过促进CD6 4的表达 ,增强体液免疫应答效应阶段单核 巨噬细胞的免疫效应功能 ,但通过抑制CD2 3的表达 ,下调IgE介导的体液免疫效应。     

豚鼠哮喘模型气道重建对气道反应性的影响

目的:通过小剂量致敏原反复致敏建立豚鼠慢性哮喘模型,设定不同时点(4、6、8周)观察气道结构及气道反应性的变化,探讨气道重建对气道反应性的影响。方法:采用低剂量卵白蛋白反复致敏、激发豚鼠,制作哮喘动物模型,应用氯化乙酰胆碱静脉注射进行支气管激发实验,进行支气管肺泡灌洗并计数灌洗液的白细胞总数及分类。应用图像分析系统对豚鼠气道进行形态学测量。结果:哮喘8周组以纤维组织增生及平滑肌厚度增加最为显著。气道高反应性以哮喘4周组增高最明显,而随着与致敏原接触时间的延长,气道高反应性逐渐降低,到哮喘8周组与正常对照组相比已无显著差异。结论:长时间卵白蛋白吸入致敏可以引起哮喘豚鼠气道反应性的变化,这种变化与气道重建的发生相关联。     

Morphological features of nerve terminal degeneration as part of the remodeling process in the motor endplate in adult muscles

With the exception of signs of retraction and withdrawal, there have been few morphological data concerning degenerated neural profiles in adult motor endplates. Here, investigation into the ultrastructure of the soleus motor endplates of adult rats (4 months old) turned up particular axonal degeneration in approximately 3% of the subjects. These axons occur as synaptic debris in the synaptic matrix of the motor endplate, adjacent to thin processes of the perisynaptic cells occupying the outer most layer of the motor endplate and were devoid of basal lamina. They often possessed dense-cored vesicles (50-80 nm). Axonal debris released from Schwann cell processes occurred during the period of acute sciatic neurectomy, when nerve terminals progressively disrupted within the motor endplate associated Schwann cells. Finally, immunohistochemical staining for antibodies to label macrophages (ED1 or ED2) has shown that nerve fiber-associated macrophages are located near the motor endplate. The results suggest that during the course of endplate remodeling, a few parts of the terminal branches are disposed of through spontaneous collapse, subsequent release from the Schwann cell investment, and eventual ingestion by macrophages in the perisynaptic space.     

Evaluation of the relationship between IL-18 levels in urine and parameters of renal pathological changes in patients with lupus nephritis]

AIM: To evaluate the relationship between IL-18 levels in urine and parameters of renal pathological changes in patients with lupus nephritis (LN). METHODS: IL-18 levels in morning free urine and 24-hour's urine in 19 normal persons and 55 patients with LN were measured by ELISA. The correlation between IL-18 levels and parameters of renal pathological changes, namely activity index (AI) and chronicity index (CI), were analyzed by liner correlation analysis method. RESULTS: IL-18 levels in morning free urine and 24-hour's urine in LN group were elevated significantly compared with control group. In both groups IL-18 levels in morning free urine were (247.1+/-317.5) ng/L and (20.3+/-14.5) ng/L, respectively, P<0.001; those in 24-hour's urine were (192.1+/-170.1) ng/d and (21.0+/-3.8) ng/d, respectively, (P<0.001). There was close positive correlation between IL-18 levels in morning free urine and 24-hour's urine and LN patient's AI (for morning free urine: r=0.602, P<0.001; for 24-hour's urine: r=0.461, P<0.005) but there was no correlation between IL-18 levels in morning urine and 24-hour's urine and CI (P>0.05). Patients with LN were divided into three groups (high, moderate and low) according to AI value. There was distinct difference of IL-18 levels in urine among the three groups: IL-18 levels in morning urine were (69.2+/-82.7) ng/L, (193.5+/-106.1) ng/L and (580.7+/-453.1) ng/L, respectively, (P<0.001); those in 24-hour's urine were (103.5+/-141.4) ng/d, (188.8+/-124.0) ng/d and (333.1+/-183.2) ng/d, respectively. CONCLUSION: It is very simple and convenient to detect IL-18 levels in morning free urine, so it is a good method for evaluating renal pathological activity of LN.     

A tetracycline-regulatable adeno-associated virus vector for double-gene transfer

An increasing demand for polycistronic vectors that express multiple genes simultaneously has arisen in recent years to obtain an efficient gene therapy. Armed with the knowledge that the expression of transgene in mammalian cells often requires tight control, we constructed in this study a tetracycline-regulated double-gene adeno-associated virus (AAV) vector carrying green and red fluorescent protein genes and expressed it in PC12 cells. When detected by fluorescence microscope and fluorescence-activated cell sorting, gene expression was induced by 44-66-fold and could be reversibly controlled by doxycycline. This double-gene AAV vector may be useful for regulated expression of two genes or a marker to monitor transgene expression.     

Protective cytotoxic T lymphocyte responses against paramyxoviruses induced by epitope-based DNA vaccines: involvement of IFN-gamma

Plasmid DNA vectors have been constructed with minigenes encoding a single cytotoxic T lymphocyte (CTL) epitope from either the M2 protein of respiratory syncytial virus (RSV) or from the nucleoprotein of measles virus (MV) with or without a signal sequence (also called secretory or leader sequence). Following intradermal immunization, plasmids in which the CTL epitopes were expressed in-frame with the signal sequence were more effective at inducing peptide- and virus- specific CTL responses than plasmids expressing CTL epitopes without the signal sequence. This immunization resulted in protection against MV-induced encephalitis and a significant reduction in viral load following RSV challenge. The reduction of viral load following RSV challenge was abrogated by prior injection with anti-IFN-gamma antibodies. These results highlight the ability of epitope-based DNA immunization to induce protective immune responses to well-defined epitopes and indicate the potential of this approach for the development of vaccines against infectious diseases.      

Identification of MEN1 gene mutations in sporadic carcinoid tumors of the lung

Lung carcinoids occur sporadically and rarely in association with multiple endocrine neoplasia type 1 (MEN1). There are no well defined genetic abnormalities known to occur in these tumors. We studied 11 sporadic lung carcinoids for loss of heterozygosity (LOH) at the locus of the MEN1 gene on chromosome 11q13, and for mutations of the MEN1 gene using dideoxy fingerprinting. Additionally, a lung carcinoid from a MEN1 patient was studied. In four of 11 (36%) sporadic tumors, both copies of the MEN1 gene were inactivated. All four tumors showed the presence of a MEN1 gene mutation and loss of the other allele. Observed mutations included a 1 bp insertion, a 1 bp deletion, a 13 bp deletion and a single nucleotide substitution affecting a donor splice site. Each mutation predicts truncation or potentially complete loss of menin. The remaining seven tumors showed neither the presence of a MEN1 gene mutation nor 11q13 LOH. The tumor from the MEN1 patient showed LOH at chromosome 11q13 and a complex germline MEN1 gene mutation. The data implicate the MEN1 gene in the pathogenesis of sporadic lung carcinoids, representing the first defined genetic alteration in these tumors.      

Development of an inhaled endotoxin challenge protocol for characterizing evoked cell surface phenotype and genomic responses of airway cells in allergic individuals.

BACKGROUND: Environmental exposure to endotoxin is a known cause of exacerbation of asthma. Inhaled endotoxin protocols have been used to evaluate airway cell surface phenotypes associated with antigen presentation and innate immunity in healthy volunteers, but not in allergic volunteers. OBJECTIVES: To establish the safety of challenge with low-dose endotoxin (10,000 endotoxin units) (lipopolysaccharide [LPS]) inhalation in allergic individuals, to measure airway cell surface phenotypes associated with antigen presentation and innate immunity in induced sputum (IS) after LPS challenge, and to conduct gene expression profiling in IS cells to determine which host genetic networks are modified by LPS inhalation. METHODS: Induced sputum was obtained before and 6 hours after LPS inhalation in 10 allergic volunteers (8 with asthma and 2 with rhinitis). Flow cytometry was used to examine cell surface phenotypes on IS cells. Genomic expression was analyzed on a subset of IS samples (n = 10) using microarray and ingenuity pathway analysis. RESULTS: A total of 10,000 endotoxin units of LPS induced significant up-regulation of membrane CD14, CD11b, CD16, HLA-DR, CD86, and Fcepsilon receptor 1 on sputum phagocytes and increased expression of genes that influence antigen-presenting surface molecules (HLA-DR, chemokine ligand 2 or monocyte chemoattractant protein 1, v-rel reticuloendotheliosis viral oncogene homolog, prostaglandin-endoperoxide synthase 2 or cyclooxygenase 2, and transforming growth factor beta), immune activation (CD14, interleukin 1beta, and regulated upon activation, normal T cell expressed and secreted), and inflammation (intracellular adhesion molecule 1 and inhibitory kappaBalpha). Gene profiles for nuclear factor kappaB, interleukin 1, and tumor necrosis factor pathways were also significantly affected. CONCLUSIONS: Low-dose inhaled endotoxin challenge is safe in allergic individuals with mild to moderate disease. It enhances airway cell surface phenotypes and expression of genes associated with antigen presentation, innate immunity, and inflammation. Microarray with ingenuity pathway analysis can be successfully applied to sputum cells to characterize genetic responses to inhaled exacerbants.     

Paramagnetic macrocyclic complexes as contrast agents for MR imaging: proton nuclear relaxation rate enhancement in aqueous solution and in rat tissues

Paramagnetic macrocyclic chelates show promise as magnetic resonance (MR) imaging contrast agents due to stability and relaxivity comparable to those of DTPA-type chelates. For the three copper and manganese macrocyclic complexes studied in aqueous solution, T1 and T2 relaxivities ranged from 0.14 to 5.88 mM-1sec-1 at 6.25 MHz. In rats, the intravenous administration of 16 mumol/kg of Mn(cyclam) caused the liver T1 relaxation rate to double at 15 minutes after injection. T1 measurements by pulsed MR imaging and manganese analyses on excised tissue showed that both relaxation rate (1/T1) and manganese content of liver and kidney increase linearly with the dosage of Mn(cyclam). The linear relationship between 1/T1 and manganese content can be considered an "in tissue" relaxivity plot for the agent. The resulting relaxivity is 54 mM-1sec-1 in liver, compared with 3.1 mM-1sec-1 in aqueous solution. Although this work is preliminary, the implication for medical MR imaging applications is that macrocyclic contrast agents can be effective at approximately one-tenth the current typical dose used for gadolinium DTPA.